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Comparison of mitogens from Mycoplasma pulmonis and Mycoplasma neurolyticum.

Studies on Mycoplasma pulmonis and Mycoplasma neurolyticum mitogenesis demonstrated that macrophages are not essential for the interactions of M. neurolyticum with B lymphocytes, whereas M. pulmonis that stimulates B and T lymphocytes depends on macrophages to fully exert its mitogenic effects. Membranes of both species carry the mitogenic as well as pathogenic potentials. However, M. neurolyticum major mitogens are biochemically distinct from those of M. pulmonis. Furthermore, M. pulmonis mitogenesis is directly correlated to the mycoplasmal-induced pathogenic effects, while no such correlation could be established with M. neurolyticum. It was, therefore, concluded that M. pulmonis and M. neurolyticum differ in the biochemical and biological natures of their mitogens.

Animals↗

Superiority of the chimpanzee animal model to study the pathogenicity of known Mycoplasma pneumoniae and reputed mycoplasma pathogens.

As far as we know, humans are the only known natural hosts for M. pneumoniae disease. Whereas volunteer studies have provided useful data on the pathogenesis of disease and efficacy of vaccines, experimentally inducing disease in humans raises serious ethical questions and has become increasingly difficult to defend. Thus, there is a genuine need for a satisfactory animal model to study M. pneumoniae disease. Using the cotton rat and developing chick embryo models, Eaton and co-workers (9-13) have clearly shown that the infectious "Eaton agent" was the cause of primary atypical pneumonia. After the causative agent was identified as M. pneumoniae (14), more definite and quantitative studies were possible. The hamster animal model has provided most of our information on the pathogenicity of strains, the pathogenesis of disease and the potency of inactivated vaccines. However, protective data obtained in hamsters immunized with the TS mutant vaccines did not correlate with data obtained in humans, raising concern regarding the use of the hamster animal model to evaluate the potency of live TS vaccines. The chimpanzee animal model has a number of advantages. Chimpanzees become clinically ill, show positive X-ray findings, and develop cold agglutinin titers. In fact, the experimentally induced disease in chimpanzees is remarkably similar to naturally occurring primary atypical pneumonia in patients. Because of the close genomic relationship, immunologic reagents prepared and used for human studies can also be used successfully in chimpanzee studies. The chimpanzee model also has some serious disadvantages. They are expensive to house and maintain and are generally not available to the scientific community. Nonetheless, chimpanzees are probably the best, most meaningful animal models established thus far to examine the infectious process, the immune response and the pathogenesis of this disease and to determine approaches to effective therapy and immunization of diseases produced by known pathogens, like M. pneumoniae, as well as reputed mycoplasma pathogens, such as M. genitalium and M. hominis.

Animals↗

Attachment of Mycoplasma salivarium and Mycoplasma orale to glass surfaces.

Mycoplasma salivarium ATCC 23064 and 24 other oral strains, and Mycoplasma orale ATCC 15539 and 22 other oral strains were examined quantitatively for attachment to glass surfaces by using ELISA method. Although all of the tested strains attached to glass surfaces, M. salivarium attached far more readily than M. orale. The results suggested that electrostatic bonds were involved in the attachment and that bivalent metal ions also played a role.

Bacterial Adhesion↗

Early phases of degeneration of mycoplasma-like bodies in plant tissue cultures infected with aster yellows: morphological analogies with Mycoplasma hominis.

Several forms of mycoplasma-like bodies (MLBs) were observed in electron micrographs of phloem cells from explants infected with aster yellows disease. The structures were found only in explants 11--25 days old, when the degenerating MLBs were first observed. They resembled those described by Anderson and Barile in 1965 for Mycoplasma hominis. A sequence of events in the degeneration of MLBs is postulated and morphological analogies with degenerating M. hominis are discussed.

Culture Techniques↗

The pathogenicity of Mycoplasma ovipneumoniae and Mycoplasma arginini in ovine and caprine tracheal organ cultures.

The effects of M. ovipneumoniae and M. arginini on tracheal organ cultures prepared from a neonatal kid and a foetal lamb were studied. Both organisms were isolated from the cultures throughout the 14 days of observation. M. ovipneumoniae produced ciliostasis and loss of cilia, confirmed by scanning electron microscope (SEM), after 4 days. These effects were sudden and profound in lamb explants, and gradual and less pronounced in kid explants. Clusters of organisms attached to epithelial surfaces and in association with cilia were visible by SEM. M. arginini also induced ciliostasis and cilia loss in both kid and lamb explants, but onset was more rapid, at 2 days, and there was evident recovery after day 6, with apparent regeneration of cilia. No clearly recognizable mycoplasmas were observed by SEM in M. arginini-infected explants.

Animals↗

Homologous regions shared by adhesin genes of Mycoplasma pneumoniae and Mycoplasma genitalium.

A lambda gt11 library of Mycoplasma genitalium genomic DNA was generated, and clones were identified using a pool of monoclonal antibodies directed against different epitopes of the 140 kDa adhesin protein. Because the 140 kDa protein of M. genitalium and the 170 kDa P1 adhesin of M. pneumoniae share biological properties such as a tip-associated location, cytadherence function and immunologic crossreactivity, we performed Southern blot analysis using these cloned partial 140 kDa gene fragments and 14 subclones that span the P1 structural gene of M. pneumoniae. Homologous regions of the two genes were identified.

Adhesins, Bacterial↗

Prevalence of Mycoplasma haemofelis, 'Candidatus Mycoplasma haemominutum', Bartonella species, Ehrlichia species, and Anaplasma phagocytophilum DNA in the blood of cats with anemia.

Hemoplasmas are known causes of anemia in some cats and some Bartonella species have been associated with anemia in people and in dogs. In this retrospective study, we used polymerase chain reaction (PCR) assays to determine the prevalence rates of Mycoplasma haemofelis, 'Candidatus M haemominutum', A phagocytophilum, Ehrlichia species, and Bartonella species DNA in the blood of cats with anemia and a control group of healthy cats. DNA of the organisms was amplified from 22 of 89 cats with anemia (24.7%) and 20 of 87 healthy cats (23.0%). DNA of a hemoplasma was amplified from 18 of 89 cats with anemia (20.2%) and 13 of 87 healthy cats (14.9%); DNA of a Bartonella species was amplified from five of 89 cats with anemia (5.6%) and seven of 87 healthy cats (8.0%). There were no statistically significant differences detected between groups.

Anaplasma phagocytophilum↗

Molecular differentiation of Mycoplasma gallisepticum and Mycoplasma imitans strains by pulsed-field gel electrophoresis and random amplified polymorphic DNA.

Pulsed-field gel electrophoresis (PFGE) and random amplified polymorphic DNA (RAPD) analysis were used to compare 21 Mycoplasma gallisepticum strains and five M. imitans strains. Each strain of M. gallisepticum typed by PFGE and RAPD methods was genetically quite unique and RAPD and PFGE fingerprinting enabled strain characterization. Relationships between the M. gallisepticum and M. imitans strains were established and dendrograms were drawn from PFGE and RAPD patterns. PFGE group A and RAPD group D were significantly associated with M. imitans strains (P < 0.05). Three M. imitans strains shared the same PFGE and RAPD patterns. The two M. gallisepticum vaccine strains had singular PFGE and RAPD patterns. Thus, PFGE and RAPD can be used to investigate disease outbreaks in vaccinated flocks or for epidemiological tracking. For M. gallisepticum, the RAPD and PFGE discriminatory powers were superior to 0.95 and the in vitro, in ovo and in vivo reproducibility of RAPD and PFGE was 100%. The RAPD drawback was the inconsistent band intensity complicating the interpretation of patterns, while the PFGE limit was its low typeability (86%). Thus, these two molecular typing methods seemed complementary for M. gallisepticum epidemiological studies.

Animals↗

Derrick Edward Award Lecture. The pathogenic potential of mycoplasmas: Mycoplasma pulmonis as a model.

For years the ubiquity of Mycoplasma pulmonis in rodents overshadowed its pathogenic potential. Its etiologic significance in murine chronic respiratory disease was established only by recognition of the delicate equilibrium between organism and host. Environmental factors and genetic predisposition of the host rather than microbial virulence are the critical determinants of disease. The prevalence of M. pulmonis is undoubtedly related to the recently demonstrated in utero transmission and the ability of the organism to colonize and produce disease in the genital tract. The etiologic significance of Ureaplasma urealyticum in human genitourinary disease, like its murine counterpart, has been surrounded by controversy. Recent studies indicate that only a subgroup of colonized individuals develop clinical manifestations of disease, ranging from infertility to fetal wastage. While the natural occurrence of both respiratory and genital mycoplasmoses seriously restricts the usefulness of rats and mice for other research purposes, they represent useful models fo the study of human disease. The recognized morphologic similarities and similar natural histories of chronic bronchitis, bronchiectasis, and emphysema in humans and of M. pulmonis respiratory disease in rats and mice make the latter a particularly useful model for study of the pathogenesis of chronic pulmonary inflammation. At the same time, murine genital mycoplasmosis represents one of the few naturally occurring genital tract diseases in laboratory animals and therefore makes an attractive model for elucidating those subtle host-parasite interactions that predispose to genital disease and subsequent reproductive failure.

Animals↗

Association of mycoplasma and human immunodeficiency virus infection: detection of amplified Mycoplasma fermentans DNA in blood.

A cross-sectional study was undertaken to determine the prevalence of Mycoplasma fermentans infection in patients with human immunodeficiency virus (HIV) infection using polymerase chain reaction methodology. Targeted M. fermentans DNA sequences could be amplified from the DNA extracted from the blood of 6 (11%) of 55 HIV-seropositive patients but from none of 26 HIV-seronegative subjects at low risk for HIV infection (P = .17). There was no correlation between M. fermentans infection and HIV clinical stage. There was a nonsignificant trend toward an association between M. fermentans infection and a history of syphilis. Infection with M. fermentans may occur more commonly in HIV-infected patients; however, a role as a copathogen or opportunistic infection was not established in this study.

Adult↗

Typing of isolates of Mycoplasma pneumoniae and Mycoplasma salivarium by growth inhibitors derived from mycoplasmal cells treated with chloroform.

Isolates of Mycoplasma pneumoniae and M. salivarium could be subclassified at the strain level by inhibitors (ch-Mcin) derived from mycoplasmal cells treated with chloroform. Sixty-one isolates of M. pneumoniae obtained from oral cavities of patients were divided into three types by their ch-Mcin: a definite type which completely inhibits the growth of M. fermentans PG18, an indefinite type and a noinhibition type. Sixty-seven isolates of M. salivarium were also divided into similar types by their ch-Mcin which does or does not inhibit the growth of M. salivarium Hup127. In the case of isolates of M. pneumoniae and M. salivarium belonging to the indefinite type which gave ambiguous patterns in the ch-Mcin typing, it was demonstrated that they could be clearly typed by further testing after repeated cloning.

Chloroform↗

TLR2 and TLR4 differentially regulate B7-1 resulting in distinct cytokine responses to the mycoplasma superantigen MAM as well as to disease induced by Mycoplasma arthritidis.

Mycoplasma arthritidis mitogen (MAM) is a superantigen secreted by M. arthritidis, an agent of murine arthritis and toxicity. We previously demonstrated that C3H mouse sub-strains differing in expression of Toll-like receptor 4 (TLR4), differed in immune reactivity to MAM due to differential engagement of TLR2 and TLR4. Here we examine the role of B7 co-stimulatory molecules in immune outcome and disease manifestations resulting from these different MAM/TLR2 and MAM/TLR4 interactions. Injections of MAM into C3H/HeJ mice upregulated expression of B7-1 but not B7-2 on peritoneal adherent cells, whereas B7-1 expression was lower on cells from C3H/HeSnJ mice. Anti-B7-1 antibody but not anti-B7-2, injected in vivo, changed the type 1 cytokines in MAM-injected C3H/HeJ mice to a type 2 cytokines and, conversely, the type 2 response in C3H/HeSnJ mice injected with anti-B7-1 shifted to a type 1 pattern. Whereas anti-B7-2 exerted no effect on disease in either mouse strain, anti-B7-1 significantly delayed the lethal toxicity of M. arthritidis in C3H/HeJ mice but enhanced arthritis in C3H/HeSnJ mice. Thus, TLR-mediated regulation of B7-1 results in diverse cytokine profiles in C3H sub-strains, and that the interaction of MAM with different TLR(s) may differentially affect cytokine responses and ultimately, M. arthritidis disease.

Animals↗

Mycoplasma pneumoniae--stimulation of lymphocytes obtained from adenoid vegetations and blood in children with and without serological evidence of Mycoplasma pneumoniae infection.

Thymidine incorporation in lymphocytes obtained from adenoids (AVL) and blood (PBL) and stimulated in vitro with Mycoplasma (M.) pneumoniae antigen, was determined in 16 children with serological indication of a current or recent M.pneumoniae infection, and in 14 controls. Both AVL and PBL gave higher responses in the M.pneumoniae-positive group than in the control group. The best discrimination between the two groups was achieved by AVL stimulated by M.pneumoniae, AVL did not respond, indicating that in this assay recent or actual M.pneumoniae infection is better expressed by AVL than by PBL.

Adenoids↗

Chronic proliferative arthritis of mice induced by Mycoplasma arthritidis: demonstration of a cell-mediated immune response to mycoplasma antigens in vitro.

Lymphocytes taken from mice chronically infected with Mycoplasma arthritidis exhibited a significant blastogenic response as measured by (3H)thymidine uptake when exposed in vitro to M. arthritidis antigens. The lymphocytes taken from 9 of 12 control mice of similar age exhibited an inhibition of (3H)thymidine uptake when exposed to M. arthritidis antigens.

Animals↗

Shared epitopes between Mycoplasma pneumoniae major adhesin protein P1 and a 140-kilodalton protein of Mycoplasma genitalium.

Previous serological data have demonstrated cross-reactive antigens between two pathogenic species of mycoplasmas, M. pneumoniae and M. genitalium. Preliminary analysis of sera and monoclonal antibodies (MAbs) to protein antigens of these species showed an immunodominance of adhesin P1 (165 kilodaltons [kDa]) of M. pneumoniae in mice and hamsters and a 140-kDa protein of M. genitalium in mice and experimentally infected chimpanzees. To further characterize these two proteins, we assayed multiple anti-P1 and anti-140-kDa protein MAbs by enzyme-linked immunosorbent assay, immunoblot, and radioimmunoprecipitation techniques. The 140-kDa M. genitalium protein was shown to be surface accessible and insensitive to levels of trypsin which readily degrade protein P1. Peptide mapping was used to identify a unique class of MAbs which bound a cross-reactive molecule common to both the major adhesin protein P1 of M. pneumoniae and the 140-kDa protein of M. genitalium. MAbs generated against both M. pneumoniae and M. genitalium which were reactive with this determinant blocked M. pneumoniae attachment to chicken erythrocytes.

Antibodies, Monoclonal↗

An ultrastructural study of avian synovium infected with an arthrotropic Mycoplasma, Mycoplasma synoviae.

Live Mycoplasma synoviae cells in agar base medium were inoculated intravenously into 10 2-week-old chickens. Joint tissues were collected over a 3-week period for light and transmission electron microscopy. In infected chickens, membrane-bound vacuoles of intact M. synoviae cells were seen in the cytoplasms of adipose cells. Cytoplasmic lipid-containing inclusions had accumulated in enlarged fibroblasts in tendon sheaths and loose connective tissue. Synovial cells had proliferated and contained increased amounts of granular endoplasmic reticulum and lipid inclusions. No abnormalities were seen in control chickens. Infected chickens were positive and control chickens were negative for rheumatoid factor activity by two serological tests: tanned cell hemagglutination and latex agglutination tests.

Animals↗

Differential serologic response to Mycoplasma ovipneumoniae and Mycoplasma arginini in lambs affected with chronic respiratory disease.

An enzyme-linked immunoabsorbent assay (ELISA) was used to evaluate the levels of antibodies to Mycoplasma ovipneumoniae and M. arginini in lambs with chronic respiratory disease. Sera were obtained from lambs in several flocks at various stages of the clinical disease and tested with sodium dodecyl sulfate (SDS)-treated M. ovipneumoniae and M. arginini whole cells and a crude capsular extract of M. ovipneumoniae as the antigens. There were low levels of antibody to M. ovipneumoniae in flocks sampled at the early stages of infection, whereas increased levels of antibody were present in lambs from flocks that had apparently recovered from the clinical disease. Slowly rising titers of circulating antibodies to M. ovipneumoniae were confirmed by sequential bleeding of lambs during the course of the clinical disease. However, antibody levels of M. arginini were more likely to increase earlier in the disease process. There was significant cross-reactivity between the 2 SDS-treated antigens in both the ELISA test and western immunoblotting. In contrast, the crude capsular extract was specific for detecting antibodies to M. ovipneumoniae.

Animals↗

Comparative pathogenicity of Mycoplasma bovis and Mycoplasma dispar for the respiratory tract of calves.

Mycoplasma bovis and M. dispar are both important in the etiology of calf respiratory disease, but marked differences are evident in the pathogenesis of disease caused by the two species. Both attach to bovine cells but vary in their invasiveness. M. bovis is capable of invading the lung parenchyma from the airways, while M. dispar predominantly infects the ciliated epithelial cells lining the respiratory tract. M. dispar causes ciliostasis in fetal-bovine tracheal organ cultures, whereas M. bovis does not have the same dramatic effect on ciliary activity. The immunological response of the host contributes to the lesions produced by M. bovis. Both T- and B-lymphocytes accumulate in the lung, which becomes a site for the production of antibody. In contrast, M. dispar produces a mild cellular response in the lungs of calves and may have immunosuppressive properties.

Animals↗